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Published on: June 13, 2022
Semirational Design of an Engineered Amine Dehydrogenase Enables Enantioselectivity Inversion for
Xiaohan Zhang1,2,3, Qingxia Luo1,2, Mengyu Li1,2
1State Key Laboratory of Food Nutrition and Safety, Key Laboratory of Industrial Fermentation Microbiology, College of Biotechnology, Tianjin University of Science and Technology, Tianjin 300453, China.
Abstract:
Using 4-phenylbutan-2-one as the model substrate, an (R)-enantiospecific AmDH (M0) derived from l-amino acid dehydrogenase was converted into (S)-enantioselective enzymes (M4-M7) through several rounds of semirational iterative mutations. Variants M6-1 and M7 were successfully applied to synthesize (S)-amines (70-81% yields) and (R)-β-amino alcohols (60-65% yields) with 95% to >99% ee. Molecular dynamics simulations provided insights into the role of mutations in substrate recognition and stereoselective control, offering important guidance for regulating the stereoselectivity of AmDHs through protein engineering.
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